JP6933659B2 - How to treat triple-negative breast cancer - Google Patents

Description

The present invention relates to a method of treating a patient diagnosed with triple negative breast cancer.

Breast cancer is the most common cancer in women. Breast cancer is a heterogeneous disease with various physiological characteristics and clinical responses. Breast cancer types are divided based on receptor expression.

There are three molecules identified as promoting many breast cancers: the estrogen receptor (ER), the progesterone receptor (PR) and the human epidermal growth factor receptor 2 (HER2).

Triple-negative breast cancer (“TNBC”) refers to any breast cancer that does not express a gene for the estrogen receptor (ER), progesterone receptor (PR) or HER2 / neu (HER2), and is about breast cancer. It accounts for 15%. Tumors that express ER and PR are treated with agents that interfere with the production or action of hormones. Tumors that have amplified HER-2 / Neu are treated with agents that inhibit HER-2 / Neu. This makes the treatment of TNBC more difficult, as most chemotherapy targets one of the three receptors. Triple-negative breast cancer has a very different pattern of recurrence than hormone-positive breast cancer, that is, the risk of recurrence is much higher during the first 3-5 years, but then drops sharply in hormone-positive breast cancer. Substantially below the risk of recurrence. Therefore, TNBC is a very aggressive and poorest prognosis type of breast cancer. This recurrence pattern is found in all types of triple-negative cancers, although absolute recurrence and survival rates vary by subtype, and sufficient data are available. TNBC is typically high grade (insufficient differentiation), progresses rapidly, and has a higher risk of recurrence and lower survival than other subtypes of breast cancer.

Conventional treatment is limited by inadequate therapeutic response, high toxicity and development of resistance. The lack of clinically meaningful results, for example with conventional systemic and combination therapies with chemotherapy, irradiation and immunotherapy, despite their ability to stimulate and proliferate tumor antigen-specific T cells. The treatment-induced T cell response is at least partially due to the inability to overcome the tumor's immune escape mechanism that limits T cell clone proliferation after such triple therapy. There may be. Many such mechanisms have been identified, including increased tumor pressure, decreased antigen presentation, loss of antigenicity, cytokines, immunosuppressive cells, and immune checkpoints.

In view of these issues, an object of the present disclosure is to provide a safe, acceptable and effective method for treating patients with TNBC. These and other objectives have been achieved in accordance with this disclosure.

According to the present disclosure, a method for treating a patient suffering from TNBC and a composition for the treatment are disclosed. According to the present disclosure, taurolidine and C-2250 can be used as neoadjuvant therapy during the perioperative period for the treatment of breast cancer, including primary and metastatic breast cancer. These compounds can be used alone and in combination with other (combination) treatments, especially hormones, and are particularly advantageous for long-term use, for example after TNBC surgery for at least 5 to 10 years. C-2250 has been shown to be particularly useful as an oral therapy for patients diagnosed with TNBC. C-2250 is effective against multiple TNBC cell lines, has a long half-life in humans, is acceptable and safe.

One aspect of the present disclosure provides a method of treating a patient suffering from triple negative breast cancer (TNBC), taurolidine, taurultam, 1 or more oxathiazine-like compounds, or treating the patient with a combination thereof, or, taurolidine , Taurultam, one or more oxathiazine-like compounds, and compounds selected from the group consisting of combinations thereof, along with one or more checkpoint inhibitors, hyperthermia, low-dose chemotherapy, and interleukin-2. A method comprising treating a patient with multiple therapies selected from the group consisting of a combination therapy of (IL-2).

In one aspect, the patient is treated with taurolidine, taurolidine, one or more oxathiazine-like compounds, or a combination thereof, administered intravenously, orally, or in combination thereof.

In one aspect, the patient is administered with taurolidine, taurolidine, one or more oxathiazine-like compounds, or a combination thereof, administered intravenously, orally, or in combination thereof before and / or during the period of surgery, and after surgery. It is treated with orally administered taurolidine, taurolidine, one or more oxathiazine-like compounds, or a combination thereof.

In one aspect, the patient is treated with C-2250 administered intravenously, orally, or a combination thereof.

In one aspect, the patient is treated with C-2250 administered intravenously, orally, or in combination thereof before and / or during the period during surgery, and C-2250 administered orally after surgery.

In one aspect, the patient is treated with one or more cyclin-dependent kinase (CDK) inhibitors in combination with taurolidine, taurolidine, one or more oxathiazine-like compounds or combinations thereof. Any CDK inhibitor known in the art can be used in combination therapy. In one embodiment, a CDK4 / 6 inhibitor can be particularly useful. For example, palbociclib (PD-0332991), ribociclib (LEE011), and / or abemaciclib (LY2835219) can be used. In other embodiments, other CDK inhibitors can be used, such as the CDK 1/5 inhibitor dynasicrib (SCH-727965).

In one aspect, the patient is treated with one or more hormonal therapeutic agents, including anastrozole, letrozole and tamoxifen, with taurolidine, taurolidine, one or more oxathiazine-like compounds, or a combination thereof. In some embodiments, if the patient is a postmenopausal patient, the patient is treated with anastrozole and / or letrozole in combination with taurolidine, taurolidine, one or more oxathiazine-like compounds, or a combination thereof. .. In some embodiments, if the patient is a premenopausal patient, the patient is treated with tamoxifen in combination with taurolidine, taurolidine, one or more oxathiazine-like compounds, or a combination thereof.

The present disclosure also discloses the following treatments: i) one or more checkpoint inhibitors; ii) hyperthermia treatment; iii) low-dose chemotherapy; iv) interleukin-2 (IL-2); and v) taurolysin, taurultam. , Oxathiazine-like compounds, and a method of treating a patient suffering from TNBC by a multidisciplinary method comprising treating the patient with a plurality of compounds selected from the group consisting of combinations thereof.

In one aspect, patients with TNBC have the following treatments: i) one or more checkpoint inhibitors; ii) hyperthermia treatment; iii) low-dose chemotherapy; iv) interleukin-2 (IL-2); v) Treatment with a compound selected from the group consisting of taurolysin, taurlutum, oxathiazine-like compounds, and combinations thereof.

In one aspect, patients with TNBC have the following treatments: i) one or more checkpoint inhibitors; ii) hyperthermia treatment; iii) low-dose chemotherapy; iv) interleukin-2 (IL-2); v) Sequentially treated with a plurality of compounds selected from the group consisting of taurolysin, taurlutum, oxathiazine-like compounds, and combinations thereof.

In one aspect, patients with TNBC have the following treatments: i) one or more checkpoint inhibitors; ii) hyperthermia treatment; iii) low-dose chemotherapy; iv) interleukin-2 (IL-2); v) Sequential treatment with a compound selected from the group consisting of taurolysin, taurlutum, oxathiazine-like compounds, and combinations thereof.

In one aspect, patients with TNBC have the following treatments: i) one or more checkpoint inhibitors; ii) hyperthermia treatment; iii) low-dose chemotherapy; iv) interleukin-2 (IL-2); v) Treatment with a compound selected from the group consisting of taurolysin, taurlutum, oxathiazine-like compounds, and combinations thereof, where at least two of the above treatments are performed simultaneously or before and after (in tandem).

In one aspect, one or more checkpoint inhibitors are administered at low doses.

In one aspect, IL-2 is administered at a biologically effective dose, a maximum tolerated dose, or a dose between the biologically effective dose and the maximum tolerated dose.

Conventional chemotherapy targets one of three receptors (estrogen receptor (ER), progesterone receptor (PR) or Her2 / neu (HER2)). In contrast, the present disclosure is an unexpectedly broad range in which taurolidine, taurultum, oxathiazine-like compounds, and combinations thereof, can kill TNBC cells as well as cancer stem cells, despite their heterogeneous nature. Being a therapeutic agent for spectrum, it involves treating TNBC with these compounds.

In the figure, ^* p <0.05, ^** p <0.01, ^*** p <0.005, ^*** p <0.001.

FIG. 5 is a graph showing the effect of various concentrations of taurolidine on the cell viability of a BT-20 cell line after 24 hours (FIG. 1A). FIG. 5 is a graph showing the effect of various concentrations of taurolidine on the cell viability of a BT-20 cell line after 48 hours (FIG. 1B). FIG. 2 is a graph showing the effect of various concentrations of C-2250 on the cell viability of the BT-20 cell line after 24 hours (FIG. 2A). FIG. 2 is a graph showing the effect of various concentrations of C-2250 on the cell viability of a BT-20 cell line after 48 hours (FIG. 2B). FIG. 5 is a graph showing the effect of various concentrations of taurolidine on cell viability of MCF-7 cell lines after 24 hours (FIG. 3A). FIG. 5 is a graph showing the effect of various concentrations of taurolidine on cell viability of MCF-7 cell lines after 48 hours (FIG. 3B). FIG. 5 is a graph showing the effect of various concentrations of C-2250 on cell viability of MCF-7 cell lines after 24 hours (FIG. 4A). FIG. 5 is a graph showing the effect of various concentrations of C-2250 on cell viability of MCF-7 cell lines after 48 hours (FIG. 4B). FIG. 5 is a graph showing the effect of various concentrations of taurolidine on the cell viability of the MDA-MB-231 cell line after 24 hours (FIG. 5A). FIG. 5B is a graph showing the effect of various concentrations of taurolidine on the cell viability of the MDA-MB-231 cell line after 48 hours (FIG. 5B). FIG. 6 is a graph showing the effect of various concentrations of C-2250 on the cell viability of the MDA-MB-231 cell line after 24 hours (FIG. 6A). FIG. 6 is a graph showing the effect of various concentrations of C-2250 on the cell viability of the MDA-MB-231 cell line after 48 hours (FIG. 6B). FIG. 6 is a graph showing the effects of various concentrations of taurolidine on necrosis and apoptosis of TNBC cells after 12 hours (FIG. 7A). It is a graph showing that high doses of taurolidine and C-2250 (5 mM) had no significant effect on peripherally derived human monocytes after 24 hours (FIG. 7B). FIG. 5 is a graph showing the effects of various concentrations of C-2250 on necrosis and apoptosis of TNBC cells after 12 hours. FIG. 5 schematically illustrates the use of taurolidine and / or C-2250 as a peri-adjuvant treatment for a subject diagnosed with TNBC.

Throughout this specification, all ranges described include all values and subranges therein, unless otherwise specified. Further, the singular form ("a" or "an") for which no quantity is specified has the meaning of "1 or more" throughout the present specification unless otherwise specified.

One aspect of the present disclosure provides a method of treating a patient suffering from triple negative breast cancer (TNBC), taurolidine, taurultam, 1 or more oxathiazine-like compounds, or treating the patient with a combination thereof, or, taurolidine , Taurultam, one or more oxathiazine-like compounds, and compounds selected from the group consisting of combinations thereof, along with one or more checkpoint inhibitors, hyperthermia, low-dose chemotherapy, and interleukin-2 (IL-2). A method comprising treating a patient with multiple therapies selected from the group consisting of combination therapies.

In one aspect, the patient is treated with taurolidine, taurolidine, one or more oxathiazine-like compounds, or a combination thereof, administered intravenously, orally, or in combination thereof.

In one aspect, the patient is administered with taurolidine, taurolidine, one or more oxathiazine-like compounds, or a combination thereof, intravenously, orally, or in combination thereof before and / or during the period of surgery, and orally after surgery. It is treated with taurolidine, taurolidine, one or more oxathiazine-like compounds, or a combination thereof administered in.

In one aspect, the patient is treated with C-2250 administered intravenously, orally, or a combination thereof.

In one aspect, the patient is treated with C-2250 administered intravenously, orally, or in combination thereof before and / or during the period during surgery, and C-2250 administered orally after surgery.

In one aspect, the patient is treated with one or more hormonal therapeutic agents, including anastrozole, letrozole and tamoxifen, with taurolidine, taurolidine, one or more oxathiazine-like compounds, or a combination thereof. In some embodiments, if the patient is a postmenopausal patient, the patient is treated with anastrozole and / or letrozole in combination with taurolidine, taurolidine, one or more oxathiazine-like compounds, or a combination thereof. .. In some embodiments, if the patient is a premenopausal patient, the patient is treated with tamoxifen in combination with taurolidine, taurolidine, one or more oxathiazine-like compounds, or a combination thereof. In certain embodiments, taurolysin, taurlutum, one or more oxathiazine-like compounds, or combinations thereof with hormonal therapeutic agents, are anti-cancer agents such as IL-2, cyclophosphamide, gemcitabine, 5-fluorouracil, paclitaxel, Includes additional combinations of cisplatin, and one or more of carboplatin.

According to a particular embodiment, the disclosure is selected from the group consisting of one or more checkpoint inhibitors, hyperthermia, interleukin-2 (IL-2), and taurolidine, taurolidine, oxathiazine-like compounds, and combinations thereof. The present invention relates to a method of treating a patient suffering from TNBC by a multidisciplinary method including treating the patient with the compound. In certain embodiments, taurolidine and / or C-2250 can be used as a peri-adjuvant treatment for a subject diagnosed with TNBC, as shown in FIG.

Oxathiazine-like compounds are described in International Application PCT / IB2015 / 059741 filed December 17, 2015, which is incorporated herein by reference in its entirety.

One aspect of the disclosure is the following treatments: i) one or more checkpoint inhibitors; ii) hyperthermia treatment; iii) low-dose chemotherapy; iv) interleukin-2 (IL-2); and v) taurolysin, It relates to a method of treating a patient suffering from TNBC by a multidisciplinary method comprising treating the patient with a plurality of compounds selected from the group consisting of taurlutam, oxathiazine-like compounds, and combinations thereof.

In one aspect, patients with TNBC have the following treatments: i) one or more checkpoint inhibitors; ii) hyperthermia treatment; iii) low-dose chemotherapy; iv) interleukin-2 (IL-2); v) Treatment with a compound selected from the group consisting of taurolysin, taurlutum, oxathiazine-like compounds, and combinations thereof.

In one aspect, patients with TNBC have the following treatments: i) one or more checkpoint inhibitors; ii) hyperthermia treatment; iii) low-dose chemotherapy; iv) interleukin-2 (IL-2); v) Sequentially treated with a plurality of compounds selected from the group consisting of taurolysin, taurlutum, oxathiazine-like compounds, and combinations thereof.

In one aspect, patients with TNBC have the following treatments: i) one or more checkpoint inhibitors; ii) hyperthermia treatment; iii) low-dose chemotherapy; iv) interleukin-2 (IL-2); v) Sequential treatment with a compound selected from the group consisting of taurolysin, taurlutum, oxathiazine-like compounds, and combinations thereof.

In one aspect, patients with TNBC have the following treatments: i) one or more checkpoint inhibitors; ii) hyperthermia treatment; iii) low-dose chemotherapy; iv) interleukin-2 (IL-2); v) Treatment with a compound selected from the group consisting of taurolysin, taurlutum, oxathiazine-like compounds, and combinations thereof, where at least two of the above treatments are performed simultaneously or before and after.

In one aspect, the patient is co-administered with IL-2 and a compound selected from the group consisting of taurolidine, taurolidine, oxathiazine-like compounds, and combinations thereof. In certain embodiments, IL-2 is co-administered with taurolidine. In certain embodiments, IL-2 is co-administered with C-2250.

In one aspect, hyperthermia treatment is performed, for example, in a temperature range of 40 ° C. to 45 ° C., such as 40 ° C., 40.5 ° C., 41 ° C., 41.5 ° C., 42 ° C., 42.5 ° C., 43 ° C., 43.5. Includes local hyperthermia at ° C., 44 ° C., 44.5 ° C. or 45 ° C., whole body hyperthermia (WBHT), or a continuous combination thereof. In certain embodiments, hyperthermia therapy comprises using a radio frequency device for selective hyperthermia therapy.

In one aspect, the methods of the present disclosure reduce or suppress the proliferative capacity of TNBC cells or cancer stem cells.

In one embodiment, one or more checkpoint inhibitors are administered at low doses. For example, during treatment, two or three different checkpoint inhibitors are administered to the patient simultaneously or sequentially. In one aspect, the one or more checkpoint inhibitors are administered at a dose below the dosage approved by the Food and Drug Administration (FDA). Checkpoint inhibitors include, but are not limited to, human programmed death receptor-1 (PD-1) blocking antibodies and human cytotoxic T lymphocyte antigen 4 (CTLA-4) blocking antibodies. Checkpoint inhibitors include, but are not limited to, nivolumab, ipilimumab, pembrolizumab, tremelimumab and pidilimumab.

As a low-dose checkpoint inhibitor, 0.1 mg / kg to 1.8 mg / kg, 0.2 mg / kg to 1.5 mg / kg once a week, once every two weeks, or once every three weeks. Examples include kg, 0.4 mg / kg to 1 mg / kg, or 0.5 mg / kg. Low doses of nivolumab are 0.1 mg / kg to 1.8 mg / kg, 0.2 mg / kg to 1.5 mg / kg, 0 once weekly, once every two weeks, or once every three weeks. .4 mg / kg to 1 mg / kg, or 0.5 mg / kg. Low dose ipilimumab is 0.1 mg / kg to 1.8 mg / kg, 0.2 mg / kg to 1.5 mg / kg, 0 once weekly, once every two weeks, or once every three weeks. .3 mg / kg to 1 mg / kg, or 0.3 mg / kg. Low doses of penbrolizumab are 0.05 mg / kg to 1.5 mg / kg, 0.1 mg / kg to 1 mg / kg, 0.2 mg once weekly, once every two weeks, or once every three weeks. / Kg to 0.8 mg / kg, 0.3 mg / kg to 0.5 mg / kg, or 0.2 mg / kg, 0.3 mg / kg or 0.4 mg / kg.

In certain embodiments, the methods of the present disclosure include follow-up monitoring by measurement of circulating tumor cells (CTCs). CTCs are a subpopulation of tumor cells that originate from the site of the primary cancer and circulate in the blood. The methods of the present disclosure include PCR, such as real-time PCR (RT-PCR), multiplex PCR, purification of RNA from blood, antibody-based capture methods (using key markers CK, EpCAM, BerEP4), and Includes various strategies for enriching, detecting, and analyzing these cells, including precision filtration. CTC counting can be incorporated before or after during any step of the methods of the present disclosure. In some embodiments, if the number of CTCs exceeds a predetermined threshold, the method comprises dosing one or more anti-cancer agents with a 1.5- to 2-fold increase. If the number of CTCs is less than a predetermined threshold, the method comprises administering 0.1 to 0.8 times the dose of one or more anti-cancer agents, or does not administer IL-2 or any of the other anti-cancer agents. ..

Chemotherapy utilizes antitumor agents to prevent cancer cells from multiplying, invading, and metastasizing. As used herein, "low-dose chemotherapy" is a therapeutic dose or less, i.e., a dose less than a conventionally acceptable dose for the treatment of cancer when the pharmaceutical compound is administered by conventional means. Refers to the administration of anticancer drug compounds. Several drugs are available for the treatment of breast cancer, including cytotoxic drugs such as doxorubicin, cyclophosphamide, methotrexate, paclitaxel, thiotepa, mitoxantrone, vincristine, or combinations thereof. Endocrine therapy can be an effective treatment if the remaining breast tissue retains endocrine sensitivity. Drugs administered for this treatment include tamoxifen, megestol acetate, aminoglutethimide, fluorymesterone, leuprolide, goserelin, and prednisone. In certain embodiments, the method comprises using an anti-cancer pharmaceutical compound that downregulates TReg cells. Such compounds include, but are not limited to, cyclophosphamide, gemcitabine, 5-fluorouracil, paclitaxel, cisplatin, and carboplatin. As low-dose chemotherapy, but are not limited ^{to, 100mg / m 2 ~800mg / m} 2, 200mg / m 2 ~500mg / m 2, or anti-tumor agent at a dose of 300 mg / ^{m 2,} for example, the administration of cyclophosphamide Can be mentioned.

In one aspect, IL-2 is administered at an effective dose, a maximum tolerated dose, or a dose between the effective dose and the maximum tolerated dose. An effective daily dose of IL-2 may include pharmaceutical dosage units contained in IL-2 in the range of 1,000,000,000,000 units (U) per ^{1 m 2 of body surface area.} Also, the dosage of IL-2 will be found to be in the range of 100,000 U to 1,000,000 U per kilogram of body weight. It will be further found that the dosage of IL-2 falls within the range of 0.1 micrograms to 100 micrograms of IL-2 per kilogram of body weight.

In certain embodiments, IL-2 is 10MioI. U. / ^M 2 ~100Mio I. U. It is administered at a dose of / m ^2. In certain embodiments, IL-2 is 18Mio I.I. U. / ^M 2 ~75Mio I. U. It is administered at a dose of / m ^2. In certain embodiments, IL-2 is a 25Mio I.I. U. / ^M 2 ~60Mio I. U. It is administered at a dose of / m ^2. In certain embodiments, IL-2 is a 54Mio I.I. U. It is administered at a dose of / m ^2.

Interleukin-2 (IL-2) is a drug that has been suggested to inhibit the growth of tumor cells. However, administration of IL-2 to patients presents a serious toxicity problem as IL-2 induces a very strong systemic inflammatory response syndrome (SIRS) response in patients. do. The toxicity of IL-2 is so severe that approximately 70% of patients cannot tolerate treatment. As used herein, "IL-2" includes natural or recombinant interleukin-2, or a biologically active derivative or substantial equivalent thereof.

Methylol transfer agents such as taurolysin and taurlutam, as described in US Pat. No. 7,892,530 (Redmond and Pfirrmann), which is part of this specification by reference in its entirety. Unexpectedly, the present invention reduces or substantially eliminates the serious toxicity and side effects of IL-2 in combination therapies to inhibit tumor metastasis in patients and to treat cancer. It was found that the efficacy of IL-2 was actually enhanced by the methylol transfer agent in the combination therapy.

（式中、ＲはＨ、メチル、エチル、プロピル（例えばイソプロピル）、ベンジル等のアルキル等である）によるオキサチアジン様化合物が本発明により利用される。 In certain embodiments, formula I:

An oxathiazine-like compound (where R is in the formula is alkyl such as H, methyl, ethyl, propyl (eg isopropyl), benzyl, etc.) is utilized in the present invention.

In certain embodiments, C-2250 (tetrahydro 1,4,5-oxathiadin-4-dioxide or 1,4,5-oxathiazine-4-dioxide) is used.

The amount of compound required depends on the tumor size. In one embodiment, the invention comprises surgically reducing tumor size and treating with one or more of the above compounds. The compounds may be administered before, during or after surgery to reduce tumors. The compounds according to the invention can be administered by any suitable method, including, but not limited to, gels, capsules, tablets, intravenously, intraperitoneally and / or directly to the tumor.

The gelling agent may contain the active compound of the present invention alone, for example 2% to 4% (eg 3%) of compound 2250, or may be administered alone or may be present alone. It may be included in combination with, or it may be for topical administration. Such gels can be used to treat skin and mouth tumors, including mouth and skin squamous epithelial cell tumors. The gel can also be used to treat cervical cancer or cervical dysplasia by administering the gel to the vagina as a suppository or by syringe. The present invention may include a combination of suppositories carrying the active compound.

Solid dosage forms for oral administration include capsules, tablets, pills, powders and granules. In such solid dosage forms, the provided compositions are at least one inert and pharmaceutically acceptable excipient and / or filler or bulking agent (eg, starch, lactose, sucrose, glucose, mannitol and silicic acid). ), Binding agents (eg carboxymethyl cellulose, alginate, gelatin, polyvinylpyrrolidinone, sucrose and gum arabic (acacia)), wetting agents (eg glycerol), disintegrants (eg agar, calcium carbonate, potato starch, tapioca starch, Alginic acid, certain silicates and sodium carbonate), solution starching agents (eg paraffin), absorption enhancers (eg quaternary ammonium compounds), wetting agents (eg cetyl alcohol and glycerol monostearate), absorption It is mixed with agents (eg kaolin and bentonite clay), and lubricants (eg starch, calcium stearate, magnesium stearate, solid polyethylene glycol, sodium lauryl sulfate), and mixtures thereof. For capsules, tablets and pills, the dosage form may include a buffer.

The compounds of the present disclosure, particularly compound 2250, have been found to be highly water soluble. In certain embodiments, PVP is not required to increase solubility. For example, a 3.2% solution of 2250 is isotonic. This is an unexpected advantage over taurolidine.

Compounds of the invention, such as compound 2250 (containing or not containing taurolidine and / or tauroltum), are particularly useful in surgical oncology because the compounds do not interfere with wound healing. Since other anti-neoplastic agents interfere with wound healing and promote suture failure, administration of other such anti-neoplastic agents must be delayed until 5 weeks after surgery. Such problems can be circumvented by compounds of the invention, such as Compound 2250, which can be administered during and immediately after surgery without the problem of wound healing or suture failure.

Similar types of solid compositions can be utilized as fillers in soft and / or hard-filled gelatin capsules that use excipients such as lactose or lactose alongside high molecular weight polyethylene glycols and the like. Solid dosage forms of tablets, dragees, capsules, pills, and granules are used with coatings and shells such as enteric coatings and other coatings well known in the field of pharmaceutical formulations. Can be prepared. These dosage forms may optionally include an opacity agent, optionally in a delayed manner, to release a composition (s) provided only in or preferentially in a particular portion of the intestinal tract. It may be. Examples of embedding compositions that can be used include polymeric substances and waxes. Similar types of solid compositions can be used as fillers in soft and hard packed gelatin capsules that use excipients such as lactose or lactose alongside high molecular weight polyethylene glycols and the like.

In certain embodiments, the capsule may include an excipient formulation comprising one or more of hydroxypropyl methylcellulose (HPMC), gelatin and fish gelatin. In certain embodiments, the capsule may comprise compound 2250 in combination with taurolidine and / or taurultum. Capsules may optionally further comprise one or more of isothiocyanates such as lycopene, ellagic acid (polyphenols), curcumin, piperine, delphinidin, resveratrol, sulforaphane, capsaicin, and piperlongmin.

The active compound of the present invention, such as compound 2250, can be combined with compounds such as gemcitabine. This combination can be used to treat cancers such as pancreatic cancer. Taurolidine and / or taurultum can also be combined with gemcitabine, for example to treat pancreatic cancer.

In some embodiments, nutritional cancer prevention and treatment products are 100 mg to 500 mg of Compound 2250 alone, or 100 mg to 500 mg of taurolysin and / or taurultam and lycopine, eg, 20 mg to 200 mg, ellag. It may contain in combination with one or more of isothiocyanates such as acid (polyphenol), curcumin, piperine (20 mg-200 mg), delphinidin, resveratrol, sulforaphane, capsaicin, and piperlongmin.

Since the above compounds do not inhibit wound healing unlike other chemotherapeutic agents, it was unexpectedly found that the compounds can be administered during and immediately after surgery.

Taurolidine, taurolidine, and oxathiazine-like compounds and their derivatives have been found to unexpectedly kill tumor stem cells, which are extremely rare and probably unknown in chemotherapeutic agents. Typical chemotherapeutic agents, when effective against tumor stem cells, are generally only effective at very high doses and are highly toxic to human patients.

Unexpectedly, it was found that lower doses of taurolidine and / or taurultum than were needed to kill the tumor cells killed the tumor stem cells.

Oxathiazine-like compounds and their derivatives have a half-life in human blood that is significantly longer than the half-lives of taurolidine and taurultum. Therefore, these compounds are not removed very quickly from the patient's bloodstream and, as a result, effectively delay the loss of drug efficacy caused by the body's clearance mechanism.

Unlike the burning sensation effect observed in patients treated with taurolidine, oxathiazine-like compounds reduce this burning sensation when applied directly to tissues.

Oxatiazine-like compounds have a combination of particularly advantageous properties, including high water solubility, versatile routes of administration, including oral and intravenous, extended stability and half-life, and significantly reduced side effects of burning sensation. Relief of burning sensation makes oxathiazine-like compounds, including 2250, highly suitable for pharmaceutical use, including oral, peripheral, intravenous, intraperitoneal, and other methods of administration in which the patient may suffer from a burning sensation. ..

Another significant advantage is that compound 2250 has a half-life of more than 24 hours in human blood, which is more than the half-life of taurolidine found to be about 30 minutes using the same test. Remarkably long.

The daily dose may be from about 0.1 g to about 100 g, for example from about 5 g to about 30 g. The daily dose may be administered in the form of a composition for oral administration. The daily dose may be administered in the form of capsules, tablets or pharmaceutically acceptable solutions. The daily dose may be administered in a form comprising compound 2250 at a concentration of about 0.01% by weight / volume to about 3% by weight / volume. The daily dose may be administered in a form comprising compound 2250 at a concentration of about 0.01 μg / ml to about 1000 μg / ml. The daily dose may be administered in a form comprising one or more solubilizers, such as polyols.

In some embodiments, the compound is administered in the composition at a concentration of about 0.01 μg / ml to about 1000 μg / ml. In some embodiments, the compound is administered in the composition at a concentration of about 1 μg / ml to about 100 μg / ml. In some embodiments, the compound is administered in the composition at a concentration of about 10 μg / ml to about 50 μg / ml. The composition may also contain from about 0.01 μg / ml to 1000 μg / ml, from about 1 μg / ml to about 100 μg / ml, or from about 10 μg / ml to about 50 μg / ml taurolidine and / or taurultum.

In some embodiments, the compound is administered in the composition at a concentration of about 0.01% to about 3%. In some embodiments, the compound is administered in the composition at a concentration of about 0.1% to about 2.5%. In some embodiments, the compound is administered in the composition at a concentration of about 1% to about 2%. The composition may also contain from about 0.01% to about 3%, from about 0.1% to about 2.5%, or from about 1% to about 2% taurolidine and / or taurolidine.

In one embodiment, the oxathiazine-like compound and its derivatives can be applied as a combination therapy with taurolidine and / or taurultum to kill tumor stem cells. According to such embodiments, the combination therapy may unexpectedly require a dosage of the drug to kill the tumor stem cells, which is less than the amount required to kill the normal tumor cells. all right.

In certain embodiments, the oxathiazine-like compound and its derivatives can be administered with vitamin D3, resulting in an increased antitumor effect of the compound.

In certain embodiments, hyperthermia therapy can be applied at the same time as Vitamin C. In certain embodiments, vitamin C is in high doses, eg 0.3 g / kg to 1 g / kg or 0.5 g / kg.

In certain embodiments, hyperthermia therapy can be applied simultaneously with doses of, for example, 500 mg to 1000 mg, 600 mg, or 800 mg of alpha lipoic acid.

In one embodiment, the taurolidine, taurolidine or oxathiadin-like compound is administered to the subject at a total daily dose of about 0.1 g to about 100 g, about 1 g to about 80 g, about 2 g to about 50 g, or about 5 g to about 30 g. Can be done.

Effective dosages of taurolidine, taurolidine or oxathiazine-like compounds are from about 0.1 mg / kg to 1000 mg / kg, preferably 150 mg / kg to 450 mg / kg per day, most preferably 300 mg / kg to 450 mg / kg per day. A dosing unit included in the range. Preferred dosages can be in the range of about 10 to 20 grams of taurolidine, taurultum or mixtures thereof per dose.

Another advantage of using taurolidine, taurolidine, or oxathiazine-like compounds is the avoidance of toxicity of such treatments, which makes conventional treatments unacceptable. The use of taurolysin, taurultam or oxathiazine-like compounds in the treatment of TNBC may result in hair loss, edema, radiation burns, loss of appetite, infections, nausea, menopause, rash, aching pain or numbness in the limbs, hearing problems, It is beneficial in avoiding the side effects of existing treatments, including loss of balance, joint pain, and swelling of the legs and feet.

In some embodiments, in parallel with the administration of the IL-2 1 day to about 10000000 unit according intravenous during treatment ^m 2 ^~40000000 unit ^{m 2,} 1 day to about once by intravenous during treatment - The 2% taurolidine solution is administered 6 times, more preferably about 2-4 times per day.

As used herein, the term "pure" refers to a pure substance that is at least about 80% free of impurities and contaminants. In some embodiments, the term "pure" refers to a pure substance that is at least about 90% free of impurities and contaminants. In certain embodiments, the term "pure" refers to a pure substance that is at least about 95% free of impurities and contaminants. In some embodiments, the term "pure" refers to a pure substance that is at least about 99% free of impurities and contaminants. In some embodiments, the term "pure" refers to a pure substance free of at least about 99.5% impurities and contaminants.

In certain embodiments, the compounds, compositions and methods of the invention include the use of micronized compounds. In some embodiments, the term "micronized" as used herein refers to a particle size in the range of about 0.005 micron to 100 micron. In certain embodiments, the term "micronized" as used herein refers to a particle size in the range of about 0.5 micron to 50 micron. In certain embodiments, the term "micronized" as used herein refers to a particle size in the range of about 1 micron to 25 microns. For example, the particle size of the drug may be about 1 micron, 5 microns, 10 microns, 15 microns, 20 microns, or 25 microns.

In certain embodiments, the compounds, compositions and methods of the invention include the use of nanoparticles. The term "nanoparticle" as used herein refers to any particle having a diameter of less than 1000 nanometers (nm). In some embodiments, the nanoparticles have a diameter of less than 300 nm. In some embodiments, the nanoparticles have a diameter of less than 100 nm. In some embodiments, the nanoparticles have a diameter of less than 50 nm, such as about 1 nm to 50 nm.

A formulation suitable for injection or infusion may contain one or more solubilizers, such as isotonic solutions such as glucose containing polyhydric alcohols, to provide a solution with increased compound concentration. Such a solution is described in European Patent No. 253662. The solution can be isotonic with Ringer's solution or Ringer's lactate solution. The concentration of the compound in such a solution may be in the range of 1 g / liter to 60 g / liter.

The compound may be in crystalline form after crystallization and / or recrystallization in, for example, alcohols, ketones, esters or combinations thereof. For example, the compounds of the invention can be crystallized and / or recrystallized from alcohols such as ethanol.

Exemplary oxathiazine-like compounds include:

When used in the form of nanoparticles, the compounds of the invention described in the claims have been found to achieve higher blood concentrations. In one embodiment, the invention comprises compound 2250 alone or in combination with taurolidine and / or taurultum. For example, the present invention comprises nanoparticles of the compound of the present invention encapsulated in a capsule.

Also in certain embodiments, the invention has, for example, the activity described herein for the above compounds, eg, at least 25%, 30%, 40%, 50%, 60%, 70%, 80%. , 90%, 95%, 100% or more of the derivatives of the above-mentioned compounds having the above-mentioned activity.

Also in certain embodiments, the present invention comprises oral compositions containing the compounds described herein, including pharmaceutically acceptable solutions of the above compounds, as well as capsules, tablets and the like containing the above compositions. Concerning the composition for administration.

In certain embodiments, the present disclosure relates to oral administration of an oxathiazine-like compound to a patient. In some embodiments, the oxathiazine-like compound is formulated into a capsule. In certain embodiments, the capsule comprises 50 mg to 1000 mg of an oxathiazine-like compound. In certain embodiments, the capsule comprises 100 mg to 500 mg of an oxathiazine-like compound. In certain embodiments, the capsule comprises 200 mg to 400 mg of an oxathiazine-like compound. In certain embodiments, the capsule comprises 250 mg to 350 mg of an oxathiazine-like compound. In certain embodiments, the oxathiazine-like compound is C-2250.

In certain embodiments, the present disclosure comprises a synergistic combination of IL-2 and C-2250, including combined drug formulations containing IL-2 and C-2250, as well as IL-2 and C-2250. , For example, relating to a method comprising administering simultaneously and / or in parallel in the form of separate dosage forms or combinations. With this combination therapy, patients are protected from the toxic side effects of IL-2 monotherapy while benefiting from the efficacy of both IL-2 and C-2250 anti-TNBC. The combination therapy provides a synergistically increased efficacy as a result of the unexpected increase in efficacy of the above treatments compared to the additive effect of IL-2 and C-2250. Moreover, the reduction in adverse effects compared to IL-2 monotherapy unexpectedly further enhances the synergistic effect. In certain embodiments, the compounds of the invention can be administered to a subject or patient by any suitable means, eg, in solution, eg, topically, systemically, such as by intravenous injection.

In one embodiment, the disclosure includes a method of killing a tumor stem cell by administering to a subject in need thereof an amount of taurolidine, taurultum or a mixture thereof effective for killing the tumor stem cell. An effective amount of taurolidine and / or taurultum to kill tumor stem cells is less than the amount of taurolidine and / or tauroltum required to kill tumor cells.

In some embodiments, taurolidine, taurultam, or a mixture thereof is administered at a concentration of about 0.01 μg / ml to about 500 μg / ml in a composition that kills tumor stem cells. In some embodiments, taurolidine, taurultam, or a mixture thereof is administered at a concentration of about 0.1 μg / ml to about 100 μg / ml in a composition that kills tumor stem cells. In some embodiments, taurolidine, taurolidine, or a mixture thereof is administered at a concentration of about 10 μg / ml to about 50 μg / ml in a composition effective for killing tumor stem cells. Taurolidine is effective in killing tumor stem cells in tissue culture in vitro at 0.01 μg / ml.

In some embodiments, taurolidine, taurolidine, or a mixture thereof is administered at a concentration of about 0.001% to about 2% in a composition that kills tumor stem cells. In some embodiments, taurolidine, taurultum, or a mixture thereof is administered at a concentration of about 0.01% to about 1.5% in a composition that kills tumor stem cells. In some embodiments, taurolidine, taurultum, or a mixture thereof is administered at a concentration of about 0.1% to about 1% in a composition that kills tumor stem cells.

In one embodiment, taurolidine, taurultum, or a mixture thereof, to kill tumor stem cells, is about 0.01 g to about 50 g, about 0.1 g to about 30 g, about 0.5 g to about 10 g, or about 1 g to about. Administer to subjects in need of it in a total daily dose of 5 g.

Dosages effective for killing tumor stem cells are about 0.01 mg / kg to 500 mg / kg, preferably 1 mg / kg to 100 mg / kg per day, most preferably 5 mg per day. It is a dosing unit included in the range of / kg to 50 mg / kg.

（式中、各Ｒは、独立してＨ、アルキル、又はアリールである）、

から選択される化合物を、腫瘍幹細胞の殺傷を必要とする被験体に投与することによる、腫瘍幹細胞を殺傷する方法を含む。かかる技術は、異なる半減期を有する少なくとも２種類の化合物を使用することにより、それによって得られる薬物動態学的効果を広げる、腫瘍幹細胞を殺傷する方法を提供する。一実施形態では、化合物２２５０は、タウロリジン及び／又はタウルルタムと組み合わせて使用され得る。 In another embodiment, the present disclosure may be used in combination with taurolidine and / or taurolidine:

(In the formula, each R is independently H, alkyl, or aryl),

A method of killing a tumor stem cell by administering a compound selected from the above to a subject in need of killing the tumor stem cell. Such techniques provide a method of killing tumor stem cells by using at least two compounds having different half-lives, thereby broadening the pharmacokinetic effects obtained thereby. In one embodiment, compound 2250 can be used in combination with taurolidine and / or taurultum.

This disclosure relates to the items described below.

Item 1. A method of treating a patient with triple negative breast cancer (TNBC), treating the patient with taurolysin, taurlutum, one or more oxathiazine-like compounds, or a combination thereof, or taurolysin, taurlutum, one or more oxathiazine. Select from the group consisting of one or more checkpoint inhibitors, hyperthermia, low-dose chemotherapy, and interleukin-2 (IL-2) combination therapy , along with compounds selected from the group consisting of similar compounds and combinations thereof. A method that involves treating a patient with multiple treatments.

Item 2. The method of item 1, wherein the patient is treated with taurolidine, taurolidine, one or more oxathiazine-like compounds, or a combination thereof, administered intravenously, orally, or a combination thereof.

Item 3. Patients receive taurolidine, taurolidine, one or more oxathiazine-like compounds, or a combination thereof, administered intravenously, orally, or in combination thereof before and / or during the period of surgery, and orally after surgery. The method according to item 1 or 2, wherein the method is treated with taurolidine, taurolidine, one or more oxathiazine-like compounds, or a combination thereof.

Item 4. The method of items 1-3, wherein the patient is treated with C-2250 administered intravenously, orally, or a combination thereof.

Item 5. Patients are treated with C-2250 administered intravenously, orally, or in combination thereof before and / or during the period of surgery, and with C-2250 administered orally after surgery, items 1-. The method according to 4.

Item 6. The method of items 1-5, wherein the oral dosage form of C-2250 is a capsule that achieves a blood concentration of Cmax of 2 μg / ml or higher when taken orally.

Item 7. The method according to items 1 to 6, wherein the oral dosage form of C-2250 is a capsule that achieves a blood concentration of Cmax of 5 μg / ml or more when ingested orally.

Item 8. The method of items 1-7, wherein the oral dosage form of C-2250 is a capsule that achieves a blood concentration of Cmax of 15 μg / ml or higher when taken orally.

Item 9. Items 1 to include treating a patient with multiple therapies, including administering to the patient an effective amount of C-2250 and IL-2 without inducing toxic side effects associated with IL-2 monotherapy. 8. The method according to 8.

Item 10. The method of item 1-9, wherein at least two treatments are performed simultaneously or before and after.

Item 11. The method of item 1-10, comprising treating a patient with all of a plurality of treatments.

Item 12. Sequentially, and applying a possible Azukasuru projecting one or more checkpoint inhibitors to the patient, and subjecting the patient to hyperthermia treatment, the low-dose chemotherapy for patients, Taurolidine, taurultam, oxathiazin-like compounds, and their The method of items 1-11, comprising co-administering IL-2 with a compound selected from the group consisting of combinations of.

Item 13. The method of item 1-12, wherein IL-2 is administered at a biologically effective dose, a maximum tolerated dose, or a dose between the biologically effective dose and the maximum tolerated dose.

Item 14. IL-2 is 25MioI. U. / ^M 2 ~60Mio I. U. The method of item 1-13, which is administered at a dose of / m ^2.

Item 15. The method of items 1-14, wherein IL-2 is co-administered with taurolidine, taurolidine, or a combination thereof.

と同時投与される、項目１〜１５に記載の方法。 Item 16. IL-2 is 1 or more and the following compounds:

The method according to item 1 to 15, which is co-administered with.

と同時投与される、項目１〜１６に記載の方法。 Item 17. IL-2 is C-2250:

The method of item 1-16, which is co-administered with.

Item 18. The method of item 1-17, wherein hyperthermia therapy is applied to the patient.

Item 19. The method of item 1-18, wherein topical hyperthermia therapy is applied to the patient.

Item 20. The method of item 1-19, wherein systemic hyperthermia (WBHT) is applied to the patient.

Item 21. The method of items 1-20, wherein a continuous combination of local hyperthermia therapy and systemic hyperthermia therapy is applied to a patient.

Item 22. The method of item 1-21, wherein the patient is administered one or more checkpoint inhibitors.

Item 23. The method of item 1-22, wherein the patient is administered with two or three checkpoint inhibitors.

Item 24. The method of item 1-23, wherein the patient is co-administered with two checkpoint inhibitors.

Items 1 to 24, wherein the checkpoint inhibitor of item 25.1 or higher is administered to the checkpoint inhibitor of item 1 or higher at a dosage lower than the dosage approved by the Food and Drug Administration (FDA). Method.

Item 1 The checkpoint inhibitor of item 26.1 or higher is not a human programmed death receptor-1 (PD-1) blocking antibody and / or a human cytotoxic T lymphocyte antigen 4 (CTLA-4) blocking antibody. 25.

The method of items 1-26, wherein the checkpoint inhibitor of item 27.1 or higher is selected from the group consisting of nivolumab, ipilimumab, pembrolizumab, tremelimumab, and pidilimumab.

The method of items 1-27, wherein the checkpoint inhibitors of item 28.1 or higher are nivolumab and ipilimumab.

Item 29.1 or higher checkpoint inhibitors are administered once a week, once every two weeks, or once every three weeks at a dose of 0.1 mg / kg to 1.8 mg / kg, item. The method according to 1-28.

Item 30.1 or higher checkpoint inhibitors are administered once a week, once every two weeks, or once every three weeks at a dose of 0.3 mg / kg to 0.6 mg / kg, item. The method according to 1-29.

Item 31. The method of item 1-30, wherein low-dose chemotherapy is applied to the patient.

Item 32. The method of items 1-31, wherein the patient is administered cyclophosphamide, gemcitabine, 5-fluorouracil, paclitaxel, cisplatin, and carboplatin.

Item 33. Low-dose ^{chemotherapy,} comprising administering an anti-tumor agent of ^{100mg / m 2 ~800mg / m 2} , The method of claim 1 to 32.

Item 34. The method of items 1-33, wherein low-dose chemotherapy comprises administering an antitumor agent effective for downregulation of TReg cells in an amount effective for downregulation but below cytotoxic levels. ..

Item 35. The method of items 1-34, wherein the plurality of therapies do not induce unacceptable side effects of conventional chemotherapy and IL-2 administration.

Item 36. The method of items 1-35, comprising administering IL-2 to the patient without inducing vascular leak syndrome in the patient.

Item 37. The method of items 1-36, comprising administering IL-2 to a patient without inducing hypotension and / or renal dysfunction in the patient.

Item 38. The method of item 1-37, wherein the Karnofsky score of the patient is improved by 20% or more after multiple treatments have been given.

Item 39. The method of items 1-38, further comprising reducing neuropathy and / or neuropathic pain in a patient.

Item 40. The method of items 1-39, wherein the patient's appetite is increased after a plurality of treatments have been given.

Item 41. The method of items 1-40, further comprising reducing insomnia in a patient.

Item 42. The method of items 1-41, further comprising reducing cachexia and / or fatigue in a patient.

Item 43. The method of item 1-42, further comprising administering the vitamin to the patient.

Item 44. The method of items 1-43, further comprising administering vitamin C to the patient.

Item 45. The method of items 1-44, further comprising administering to the patient alpha lipoic acid.

Item 46. The method of items 1-45, comprising administering to the patient IL-2 in combination with taurolidine for 5-7 days.

Item 47. The method of items 1-46, comprising administering to the patient a PD-1 checkpoint inhibitor and a CTLA-4 checkpoint inhibitor once a week.

Item 48. The method of item 1-47, wherein the PD-1 checkpoint inhibitor and the CTLA-4 checkpoint inhibitor are administered to the patient for 3 to 5 weeks.

Item 49. The method of items 1-48, comprising administering to the patient a PD-1 checkpoint inhibitor and a CTLA-4 checkpoint inhibitor, and treating the patient with hyperthermia therapy in parallel.

Item 50. The method of items 1-49, wherein hyperthermia therapy is applied to the patient for 3-5 weeks.

Item 51. The method of items 1-50, wherein the hyperthermia treatment is a topical hyperthermia treatment.

Item 52. The method of items 1-51, comprising applying low-dose chemotherapy to a patient and treating the patient with hyperthermia therapy in parallel.

Item 53. The method according to item 1-52, wherein the hyperthermia treatment is a systemic hyperthermia treatment.

Item 54. The method of item 1-53, wherein the patient is treated with one or more hormonal therapies in combination with taurolidine, taurolidine, one or more oxathiazine-like compounds, or a combination thereof.

Item 55. The method of items 1-54, wherein the hormonal therapeutic agent is anastrozole, letrozole, and / or tamoxifen.

Item 56. The method of item 1-55, wherein the patient is postmenopausal and is treated with anastrozole and / or letrozole in combination with taurolidine, taurolidine, one or more oxathiazine-like compounds, or a combination thereof.

Item 57. The method of item 1-56, wherein the patient is postmenopausal and the patient is treated with tamoxifen in combination with taurolidine, taurolidine, one or more oxathiazine-like compounds, or a combination thereof.

Item 58. The method of item 1-57, wherein the combination of the hormonal therapy agent with taurolidine, taurolidine, one or more oxathiazine-like compounds, or a combination thereof further comprises administering an anticancer agent.

Item 59. The method of item 1-58, wherein the anti-cancer agent is one or more of IL-2, cyclophosphamide, gemcitabine, 5-fluorouracil, paclitaxel, cisplatin, and carboplatin.

Item 60. The method of items 1-59, wherein the oxathiazine-like compound is C-2250.

Item 61. The method of items 1-60, wherein the patient is administered one or more cyclin-dependent kinase (CDK) inhibitors.

Item 62. The method of items 1-61, wherein the CDK inhibitor is a CDK4 / 6 inhibitor.

Item 63. The method of items 1-61, wherein the CDK inhibitor is palbociclib, ribociclib, abemaciclib, dynacyclib, or a combination thereof.

Example 1: Study of Survival Rate of Breast Cancer Cells Introduction Based on the recognition of taurolysin and C-2250 as potent anti-neoplastic agents, these agents were used in the following breast cancer cell lines: BT-20 (primary breast cancer, HR). Negative), MDA-MB-231 (metastatic, HR negative) and MCF-7 (metastatic, HR positive) were tested. BT-20 and MDA-MB-231 are TNBC cell lines, but MCF-7 is not a TNBC cell line.

Materials and Methods BT-20 cell lines were cultured in EMEM supplemented with 10% fetal bovine serum (FBS) and 1% penicillin-streptomycin (pen-strept). MCF-7 was cultured in DMEM supplemented with 10% FBS, 1% pen-strep and 0.1% insulin. MDA-MB-231 was cultured in DMEM containing 10% FBS and 1% pen-strep. All cells were cultured at 37 ° C. in a humid atmosphere of _{5% CO 2.}

Chemicals: A 2% solution of taurolidine and a C-2250 ultra-high purity powder were provided by Geistlich Pharma AG of Wolhusen, the assignee of the invention. A C-2250 solution was prepared by dissolving C-2250 in PBS solution to a final concentration. The reagents were further diluted to 5 different concentrations containing taurolidine of 10 μΜ, 100 μΜ, 250 μΜ, 500 μΜ and 1000 μΜ, while C-2250 was diluted to 100 μΜ, 250 μΜ, 500 μΜ, 750 μΜ and 1000 μΜ.

Colony formation assay: The breast cancer cell lines 100 [mu] l, each optical density (BT-20 and MDA-MB-231: 1.0 × 10 5 cells /Ml;BT-474:5.0×10 ⁵ cells / ml; And MCF-7: 0.5 × 10 ⁵ cells / ml) and sowed in a 96-well plate. The cells were incubated under culture conditions for 12 hours to allow the cells to adhere. The cell medium is then removed and fresh culture medium alone (control) or various concentrations of taurolidine (10 μΜ, 100 μΜ, 250 μΜ, 500 μΜ, and 1000 μΜ) and C-2250 (100 μΜ, 250 μΜ, 500 μΜ, 750 μΜ, and 1000 μΜ). ) Was added.

Cell viability assay: After 24 hours, 10 μl of MTT solution was added to each well and the incubation was extended for an additional 2 hours. The supernatant was then removed, washed with 100 μl PBS, and then 100 μl DMSO was added. Absorbance was measured using a Microplate Plate Reader (Dynex Technologies Inc., Chantilly, Virginia) at a wavelength of 570 nm. In addition, the experiment was carried out for 48 hours, and the whole procedure was repeated 3 times.

The resulting data are expressed as mean ± SEM and are representative of three separate experiments.

The results of cell survival studies on the cell viability of the taurolidine BT-20 cell line after 24 and 48 hours are shown in FIGS. 1A and 1B.

The results of cell survival studies on the cell viability of the BT-20 cell line of C-2250 after 24 hours and 48 hours are shown in FIGS. 2A and 2B.

The results of cell survival studies on the cell viability of the taurolidine MCF-7 cell line after 24 and 48 hours are shown in FIGS. 3A and 3B.

The results of cell survival studies on the cell viability of the MCF-7 cell line of C-2250 after 24 hours and 48 hours are shown in FIGS. 4A and 4B.

The results of cell survival studies on the cell viability of the taurolidine MDA-MB-231 cell line after 24 hours and 48 hours are shown in FIGS. 5A and 5B.

The results of cell survival studies on the cell viability of the C-2250 MDA-MB-231 cell line after 24 and 48 hours are shown in FIGS. 6A and 6B.

Summary Taurolidine and C-2250 exhibit potent in vitro anti-neoplastic activity against TNBC strains. 250 μM (34 μg / ml) of C-2250 induced more than 50% cell death relative to the TNBC cell line after 48 hours. 100 μM (28 μg / ml) taurolidine induced more than 50% cell death relative to the TNBC cell line after 48 hours.

Example 2: Human Therapeutic Case Study Pre-study patient diagnosis and history:
September 1st: Histological diagnosis of triple-negative breast cancer in the right chest 3 cm, G3, CS axilla (G3, ER, PR and Her 2-new negative with histologically medullary shares) A 50-year-old white woman with. Disseminated lung metastasis.

Ki-67: 61% show very high breast cancer cell division.

Additional evidence of ductal carcinoma in situ (DCIS) with local vascular infiltration; BRCA1 negative.

Prior to entering the study, the patients were treated with a number of oncology and chemotherapy treatments, including weekly neoadjuvant taxol chemotherapy, wide local excision mammary tumor resection, and adjuvant radiation therapy for the right chest wall. rice field. These treatments were ineffective and the patient developed metastatic dissemination and neutropenia and was a terminally ill (stage IV) patient.

Research protocol:
The above patients had a Karnovsky score of 80%, mild peripherally emphasized neuropathy in all limbs, and severe pain in the left lateral chest wall during inspiration, eg, very pain, at the start of the study. He suffered from sneezing, severe shortness of breath (SOS), loss of appetite, insomnia and extreme fatigue.

diagnose:
Breast cancer ICD10: C50.9
Lung cancer ICD10: C78.6
Lymphadenopathy ICD10: R59.1

The above patients underwent a 4-week treatment protocol as follows:
Weekly low-dose checkpoint inhibitor therapy with PD1 / PDL-1 inhibition with nivolumab (0.5 mg / kg) and CTL-4 inhibition with ipilimumab (0.3 mg / kg) for 1.3 weeks,
A high frequency field (13.) using a Syncrotherm device three times per week for the chest region in combination with high doses of vitamin C (0.5 g / kg) and alpha lipoic acid (600 mg) over 2.3 weeks. Local region hyperthermia by 56 MHz), followed by
3. 3. Long-term (6-8 hours) fever range (approximately 42 ° C.) systemic hyperthermia in combination with low-dose chemotherapy using cyclophosphamide 300 mg / m ^{2 for downregulation of TReg cells, followed by}
4. Taurolidine (2% Taurolidine, intravenous 250ml) high-dose ^IL-2 (54Mio I.U./m 2 tapering regime) with combined 5 days therapy.

We have found that the combination of IL-2 and taurolidine is effective in inducing remission in TNBC and its metastases in patients who have failed with other treatments.

The day after the multidisciplinary therapy was completed, a chest x-ray demonstrated more advanced bilateral lung metastases. Induction of cytotoxic T cell responses after immunotherapy can take from 1 to 2 months up to 6 months. Therefore, two months later, the patient underwent a second chest x-ray, demonstrating a 20% improvement in reducing lung metastases as early as possible. A chest x-ray after an additional 2 months demonstrated significant partial remission.

In addition, the patient showed excellent clinical symptoms with a 100% Karnofsky score and no shortness of breath or any other cancer-related symptoms.

Both types of checkpoint inhibitors (inhibition of PD1 / PDL-1 and CTL-4) were used to avoid autoimmunity after checkpoint inhibitor therapy. Specifically, nivolumab and ipilimumab were used at lower doses that were not applicable, and were used in a regular, higher dose (weekly) order.

Discussion Low-dose cyclophosphamide or gemcitabine therapy can selectively deplete regulatory T cells (Tregs). For the success of immune-based therapies, chemotherapeutic agents (and radiation) may be combined to disrupt immune tolerance and create a tumor microenvironment. Long-term systemic hyperthermia in the mild thermal range (40 ° C-42 ° C) reduces interstitial pressure in the tumor microenvironment. In addition, hyperthermia improves the immunogenicity of cancer cells and trafficking of lymphocytes. High doses of IL-2 are known to have serious side effects, and as a result, the use of IL-2 has never been widespread. The main side effects of high-dose IL-2 therapy are induced by vascular leak syndrome, which is characterized by weight gain, anasarca, hypotension, and renal dysfunction. Vascular leak syndrome (VLS) is a life-threatening toxicity induced during IL-2 treatment in cancer patients. Co-administration of taurolidine, taurolidine, oxathiazine-like compounds, and combinations thereof reduce the side effects induced by these vascular leaks.

In summary, the disclosure describes a demonstration of the safety and efficacy of the combination of IL-2 and taurolidine in inducing remission in TNBC and its metastases in patients who have failed other treatments.

Example 3: Efficacy of Breast Cancer Cell Lines Methods: Primary and metastatic ER-negative breast cancer cells (BT-20 and MDA-MB-231) and ER-positive cells (BT-), as shown in FIGS. 7A and 8A. 474 and MCF-7) were treated separately with increasing concentrations of taurolysin and 2250. Annexin V and propidium iodide (PI) were used to assess apoptosis and necrosis based on phosphatidylserine externalization. The contribution of caspases 3, 8 and 9 to apoptosis and autophagy was evaluated by both flow cytometry and Western blotting. The effects of reactive oxygen species (ROS) were evaluated using n-acetylcysteine (NAC), a radical scavenger agent, and butionine-sulfoximine (BSO), a glutathione depleting agent. .. Necroptosis was evaluated using necrostatin-1, a potent inhibitor of the process.

RESULTS: A dose-dependent increase in cell death was observed in ER-positive and ER-negative breast cancer cells treated separately with taurolidine and 2250. Concentrations of taurolidine greater than 100 μΜ (≥284.7 micrograms) and 2250 greater than 250 μΜ (≥342 micrograms) induced more than 50% cell death after 48 hours (p <0.05-0.01). .. Cell death was most pronounced in TNBC cells. There was no evidence of autophagy involvement. Taurolidine and 2250 also induced the production of ROS, which was substantially reduced by the radical scavenger NAC and significantly increased by the glutathione depleting agent BSO.

Example 4: Safety of Taurolysin and C-2250 As shown in FIG. 7B, human peripheral blood mononuclear cells are treated with taurolysin and 2250 at concentrations ranging from 100 micromoles up to 5000 micromoles and any prominent cells. It demonstrated the safety of taurolysin and 2250 in non-neoplastic cells without inducing death.

Example 5: Half-life of 2250 A half-life of 2250 was measured at 37 ° C. in fresh human blood.

The half-life was calculated to be approximately 40 hours.

As used herein, the terms "about" and "approximately" should be determined to include any value within 5% of the listed values. In addition, the enumeration of "about" and "approximately" terms for a range of values should be determined to include both the upper and lower limits of the enumerated range. As used herein, terms such as "first," "second," and "third" uniquely identify an element and mean any particular order of the element or process. It should be judged that neither is it, nor is it limited to them.

Concentrations, quantities and other numerical data may be presented herein in a range format (eg, about 5% to about 20%). Such range formats are used for convenience and brevity only, and not only include the numbers explicitly listed as range limitations, but each number and subrange is explicit unless otherwise indicated. It is understood that it should be flexibly determined to include all individual numbers or subranges contained within that range, as listed in. For example, values in the range of about 5% to about 20%, such as, but not limited to, 5%, 5.5%, 9.7%, 10.3%, 15%, and, but not limited to, 5% to 10%. It should be determined that a partial range such as%, 10% to 15%, 8.9% to 18.9%, etc. is included.

Although the present invention has been shown and described only in some embodiments thereof, the present invention is not so limited and is subject to various modifications without departing from the spirit and scope of the present invention. It will be clear to those skilled in the art that it can be done. Further, it is understood that the embodiments shown and described of the present invention are currently regarded as preferred embodiments. Various modifications and modifications can be made to any processing step, as will be apparent to those skilled in the art who have the benefit of the present disclosure. The appended claims are to be construed as inclusive of all such modifications and modifications, and therefore the specification is intended to be regarded as a commentary rather than a limiting one. The present disclosure, including any easily identifiable variant of the teachings herein, defines, in part, the scope of the aforementioned claims so that no subject matter of the invention is devoted to the public. do. In addition, the appended claims are intended to be construed to include alternative embodiments.

Claims (16)

C-2250: For the manufacture of drugs to treat patients with triple-negative breast cancer (TNBC):

Is the use of
Use in which the agent is administered intravenously, orally, or in combination thereof . The drugs are taurolidine, taurultam,

, Or the use according to claim 1, which is used in combination with these combinations. Before Symbol agent is interleukin -2 (IL-2), 1 or more checkpoint inhibitors, for treating a patient in combination with a therapeutic of performing a plurality of combination therapy selected from hyperthermia and low dose chemotherapy The use according to claim 1 or 2. The oral dosage form of C-2250 is a capsule that achieves a blood concentration of Cmax of 2 μg / mL or more, preferably 5 μg / mL or more, more preferably 15 μg / mL or more when taken orally. Use according to any one of claims 1 to 3. The use of claim 3 , wherein at least two of interleukin-2 (IL-2), one or more checkpoint inhibitors, hyperthermia, and low-dose chemotherapy are given simultaneously or before and after. The use according to claim 3 , comprising treating the patient with interleukin-2 (IL-2), one or more checkpoint inhibitors, hyperthermia, and low-dose chemotherapy. The use according to claim 3 , wherein administration of one or more checkpoint inhibitors, hyperthermia, low-dose chemotherapy, and simultaneous administration of the drug and IL-2 are sequentially performed. IL-2 is 25000000I. U. / ^{M 2 to} 60000000 I. U. The use according to claim 3 , which is administered at a dose of / m ^2. The use according to claim 3 , wherein IL-2 is co-administered with the agent. The use according to any one of claims 3 to 9 , wherein one or more checkpoint inhibitors, preferably two or three types of checkpoint inhibitors, are administered to the patient. The one or more checkpoint inhibitor, human programmed death receptor -1 (PD-1) blocking antibody and / or human cytotoxic T-lymphocyte antigen 4 (CTLA-4) is not a blocking antibody of claim 1 0 Use as described in. The use according to any one of claims 1 to 11, wherein the patient is administered cyclophosphamide, gemcitabine, 5-fluorouracil, paclitaxel, cisplatin, and carboplatin. Wherein the agent is used in combination with one or more hormonal therapeutic agents, preferably, the hormonal therapy agent is anastrozole, letrozole and / or tamoxifen, according to any one of claims 1 to 1 2 use. It said agent, IL-2, cyclophosphamide, gemcitabine, 5-fluorouracil, paclitaxel is combined cisplatin, and with one or more carboplatin Use according to any one of claims 1 to 1 3. Wherein the agent is used in combination with one or more cyclin dependent kinase (CDK) inhibitor, preferably, the CDK inhibitor is Paruboshikuribu, Riboshikuribu, Abemashikuribu, Dinashikuribu, or CDK4 / 6 inhibitors such as combinations thereof, use according to any one of claims 1 to 1 4. C-2250 for use in methods of treating patients with triple-negative breast cancer (TNBC):

It is a drug containing
A drug in which the drug is administered intravenously, orally, or in combination thereof.

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